Substrate-attached materials are enriched with tetraspanins and are analogous to the structures associated with rear-end retraction in migrating cells
| Autor: | Tetsuji Naka, Yoshiko Yagi, Satoshi Serada, Gabriele Mugnai, Masashi Yamada, Kiyotoshi Sekiguchi |
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| Rok vydání: | 2013 |
| Předmět: |
Dynamins
Integrins cell migration Tetraspanins integrin Integrin Tetraspanin 24 Biology Tetraspanin 29 Tetraspanin 28 Focal adhesion Cellular and Molecular Neuroscience chemistry.chemical_compound proteomics Cell Movement Cell Line Tumor Myosin Cell Adhesion Humans focal adhesion Cell adhesion Dynamin Myosin Type II Focal Adhesions rho-Associated Kinases Membrane Glycoproteins ganglioside Cell migration Cell Biology CD9 Transmembrane protein Cell biology EGTA chemistry Focal Adhesion Protein-Tyrosine Kinases biology.protein Research Paper |
| Zdroj: | Cell Adhesion & Migration |
| ISSN: | 1933-6926 1933-6918 |
| DOI: | 10.4161/cam.25041 |
| Popis: | Substrate-attached materials (SAMs) are cellular feet that remain on substrates after the treatment of adherent cells with EGTA. SAMs are thought to contain cell adhesion machineries, but their biochemical properties have not been addressed in detail. To gain insight into the molecular mechanisms operating in cell adhesions, we comprehensively identified the protein components of SAMs by liquid chromatography coupled with tandem mass spectrometry, followed by immunoblot analysis. We found that the tetraspanins CD9, CD81, and CD151 were enriched in SAMs along with other transmembrane proteins that are known to associate with tetraspanins. Notably, integrins were detected in SAMs, but the components of focal adhesions were scarcely detected. These observations are reminiscent of the “footprints” that remain on substrates when the retraction fibers at the rear of migrating cells are released, because such footprints have been reported to contain tetraspanins and integrins but not focal adhesion proteins. In support of this hypothesis, the formation of SAMs was attenuated by inhibitors of ROCK, myosin II and dynamin, all of which are known to participate in rear-end retraction in migrating cells. Furthermore, SAMs left on collagen-coated substrates were found by electron microscopy to be fewer and thinner than those on laminin-coated substrates, reflecting the thin and fragile retraction fibers of cells migrating on collagen. Collectively, these results indicate that SAMs closely resemble the footprints and retraction fibers of migrating cells in their protein components, and that they are yielded by similar mechanisms. |
| Databáze: | OpenAIRE |
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